Heparin-like compounds, their preparation and use to prevent arterial thrombsis associated with vascular injury and intervetions

Abstract

The present invention is related to heparin-like compounds characterized by their capacity of inhibiting collagen-induced platelet aggregation in flowing whole blood and their use for prophylactic treatment of arterial thrombosis associated with vascular or microvascular injury and interventions. Said properties are related to a high coupling density of negatively charged heparin or heparin-like glycosaminoglycan molecules, present in multiple heparin or heparin-like glycosaminoglycans as well as in proteoglycans containing said multiple heparin or heparin-like glycosaminoglycans or lower-molecular-weight heparin or heparin-like glycosaminoglycans connected directly or through spacer/linker molecules to globular core molecules. Heparin-like compounds, with said properties are obtainable from mammalian mast cells, by tissue extraction or cell cultivation. The heparin-like compounds of the present invention can also be produced by synthetical, semisynthetical and/or biotechnological methods and they are useful for manufacturing preparations, means and devices for local or topical application in prophylactic treatment of arterial thrombosis and its sequelae.

1 . Heparin-like compounds, characterized in that they comprise heparin or heparin-like glycosaminoglycans as such, as proteoglycans and/or coupled to core molecules, said heparin-like compounds having a high coupling density of negatively charged heparin or heparin-like glycosaminoglycans either as such or connected to core molecules and said heparin-like compounds further being characterized by their capacity of substantially complete inhibition of the platelet aggregation upon collagen in flowing blood, a cause of arterial thrombosis associated with vascular or microvascular injury and interventions.  
     
     
         2 . The heparin-like compounds according to  claim 1 , characterized in that the heparin or heparin-like glycosaminoglycans have a multiple structure and comprise several end-to-end- and/or end-to-side-connected heparin or heparin-like glycosaminoglycan molecules.  
     
     
         3 . The heparin-like compounds according to  claim 1 , characterized in that the heparin or heparin-like glycosaminoglycans comprise several end-to-end-connected heparin or heparin-like glycosaminoglycan molecules.  
     
     
         4 . The heparin-like compounds according to  claim 1 , characterized in that the heparin or heparin-like glycosaminoglycans comprise several end-to-side-connected heparin or heparin-like glycosaminoglycan molecules.  
     
     
         5 . The heparin-like compounds according to  claim 1 , characterized in that the molecular weight of the heparin or heparin-like glycosaminoglycans is at least 75±25 kDa.  
     
     
         6 . The heparin-like compounds according to  claim 1 , characterized in that the molecular weight of the heparin or heparin-like glycosaminoglycans is more than 75±25 kDa.  
     
     
         7 . The heparin-like compounds according to  claim 1 , characterized in that the heparin or heparin-like proteoglycans comprise multiple heparin or heparin-like glycosaminoglycans.  
     
     
         8 . The heparin-like compounds according to  claim 1 , characterized in that they comprise multiple heparin or heparin-like glycosaminoglycans connected directly or through spacer/linker molecules to chain-like or a globular core molecules.  
     
     
         9 . The heparin-like compounds according to  claim 1 , characterized in that they comprise lower-molecular-weight heparin or heparin-like glycosaminoglycans connected directly or through spacer/linker molecules to globular core molecules.  
     
     
         10 . The heparin-like compounds according to  claim 1 , characterized in that the core molecules are of natural, synthetic or of semisynthetic origin.  
     
     
         11 . The heparin-like compounds according to  claim 1 , characterized in that the core molecule comprises a protein or a polypeptide.  
     
     
         12 . The heparin-like compounds according to  claim 1 , characterized in that the core molecule is serum albumin.  
     
     
         13 . The heparin-like compounds according to  claim 1 , characterized in that the core molecule is a polypeptide comprising one or more Ser-Gly-Ser-Gly sequences.  
     
     
         14 . The heparin-like compounds according to  claim 1 , characterized in that the heparin or heparin-like proteoglycan is mast cell derived heparin-glycosaminoglycan (HEP-PG).  
     
     
         15 . A method for producing the heparin-like compounds according to  claim 1  claim, characterized by the steps of: 
 (a) cultivating isolated and purified mast cells in a cell culture medium using conditions allowing cell proliferation;  
 (b) releasing heparin proteoglycan-containing granules by optional activation and/or lysis;  
 (c) allowing the released granules to solubilize in the surrounding culture medium;  
 (d) collecting said solubilized heparin proteoglycan (HEP-PG) from said medium;  
 (e) optionally separating the multiple heparin glycosaminoglycan (HEP-GAG) molecules from the HEP-PG-molecules.  
 
     
     
         16 . The method according to  claim 15 , characterized in that the structure of the multiple heparin glycosaminoglycans obtained in step (e) are modified by synthetic, semisynthetic or biotechnological methods.  
     
     
         17 . The method according to  claim 15 , characterized in that the activation of step (b) is carried out with mast cell agonists, which induce mast cell degranulation and release of mast cell derived HEP-PG- and HEP-GAG-molecules obtainable thereof.  
     
     
         18 . The method according to  claim 17 , characterized in that the agonist is selected from a group consisting of basic polyamines and calcium ionophores.  
     
     
         19 . A method for producing the heparin-like compounds according to  claim 1 , characterized in that several heparin or heparin-like glycosaminoglycan units are connected end-to-end and/or end-to-side by covalent binding to provide multiple heparin or heparin-like glycosaminoglycan-molecules having a high molecular weight of more than 75±25.  
     
     
         20 . A method for producing heparin-like compounds according to  claim 1 , characterized in that multiple heparin or heparin-like glycosaminoglycan molecules are connected directly or through spacer/linker molecules to core molecules.  
     
     
         21 . A method for producing heparin-like compounds according to  claim 1 , characterized in that lower-molecular-weight heparin or heparin-like glycosaminoglycan units are connected directly or through spacer/linker molecules to globular core molecules.  
     
     
         22 . A method for prophylactic treatment of arterial thrombosis associated with vascular or microvascular injuries and/or interventions, characterized by the local administration of the heparin-like compounds according to  claim 1 .  
     
     
         23 . The method according to  claim 22 , characterized in that the local administration is performed by applying an effective amount of the heparin-like compounds according to  claim 1  as such, as a preparation or in form of a coated device.  
     
     
         24 . A method for preventing interactions of flowing blood with collage and consequent platelet aggregation or clogging of blood vessels at the site of vascular injury and/or intervention, characterized in that preparations containing and/or devices coated with heparin-like compounds according to  claim 1  are administered to the site of vascular injury and/or intervention.  
     
     
         25 . Devices for local administration in connection with vascular or microvascular injuries and/or interventions, characterized in that they comprise devices coated with heparin-like compounds according to  claim 1 .  
     
     
         26 . The devices according to  claim 25 , characterized in that they comprise stents, vascular grafts or extracorporeal circulation systems.  
     
     
         27 . Preparations for local administration in connection with vascular or microvascular injuries and/or interventions, characterized in that they comprise the heparin-like compounds according to  claim 1  in combination with pharmaceutically acceptable and compatible carriers and/or adjuvants.  
     
     
         28 . A method for manufacturing devices for local administration in connection with vascular or microvascular injuries or interventions characterised in that the means or devices are allowed to be in contact with a solution containing the heparin-like compounds according to  claim 1 .  
     
     
         29 . The use of heparin-like compounds according to claims  1 , characterised in that the compounds are used for manufacturing preparations and/or devices capable of substantially complete inhibition of platelet aggregation upon collagen in flowing whole blood, the cause of arterial thrombosis associated with vascular or microvascular injury and interventions.